The Cold Spring Harbor Laboratory proposes to continue a course entitled Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging, to be held in the Fall of 2010-2014. This is a short, two week, intensive course which trains students (ranging from graduate students to Principal Investigators) to directly use advanced and/or specialized techniques in microscopy to localize nucleic acids and proteins and provides background on the biochemical concepts underlying these techniques. Students gain first-hand experience in localizing genes, chromosomes, mRNAs and proteins, by immunocytochemistry and fluorescence in situ hybridization (FISH), preparing and using labeled nucleic acid probes for FISH and labeled antibodies for immunolocalization, performing experiments using different antigen-antibody systems (fluorescence, enzymatic, colloidal gold), and employing GFP-fusion proteins to localize, quantify, and analyze proteins in living cells using standard fluorescence, fluorescence recovery after photobleaching, and resonance energy transfer. All experiments are performed using a range of state-of-the-art microscope methods (epifluorescence, confocal laser scanning, 3D deconvolution , photo-activation localization, and structured illumination) and students have the opportunity to compare the use of different systems in different applications. The techniques will be used to characterize copy number changes of oncogenes and tumor suppressor genes on the DNA-level using spectral karyotyping and comparative genomic hybridization (CGH), and to analyze the cellular distribution and the expression status of mRNAs and proteins relevant to human carcinogenesis (such as proliferation markers and tumor suppressor gene products) as well as to the pathogenesis of other human diseases in fixed and living cells. The course is taught by a team of internationally recognized leaders in the field and a team of dedicated, technically adept assistants who together provide hands-on training. Lecturers are invited who give up-to-the-minute reports that provide examples of the use of techniques the students are learning in current research.
The primary objectives of the course is to train students, postdoctoral fellows and Principal Investigators to enter directly into research that makes use of advanced and/or specialized techniques in microscopy to localize genes and gene products, as well as to provide background on the biochemical concepts underlying these techniques. Advanced techniques are becoming crucial in determining and analyzing the changes that occur in cancer cells at different stages of tumor formation and progression.
